Methods for inducing a population of T cells to proliferate using agents which recognize TCR/CD3 and ligands which stimulate an accessory molecule on the surface of the T cells

Abstract

Methods for inducing a population of T cells to proliferate by activating the population of T cells and stimulating an accessory molecule on the surface of the T cells with a ligand which binds the accessory molecule are described. T cell proliferation occurs in the absence of exogenous growth factors or accessory cells. T cell activation is accomplished by stimulating the T cell receptor (TCR)/CD3 complex or the CD2 surface protein. To induce proliferation of an activated population T cells, an accessory molecule on the surface of the T cells, such as CD28, is stimulated with a ligand which binds the accessory molecule. The T cell population expanded by the method of the invention can be genetically transduced and used for immunotherapy or can be used in methods of diagnosis.

What is claimed is:  
     
       1. A method for inducing a population of T cells to proliferate to sufficient numbers for use in therapy; comprising: 
       a) activating the population of T cells by contacting the T cells in vitro with an anti-CD3 antibody which is immobilized on a solid phase surface; and  
       b) stimulating an accessory molecule on the surface of the T cells in vitro with an anti-CD28 antibody, wherein said anti-CD28 antibody is immobilized on the same solid phase surface as the anti-CD3 antibody, the activating and stimulating steps thereby inducing proliferation of the T cells to sufficient numbers for use in therapy.  
     
     
       2. The method of  claim 1 , wherein the anti-CD3 antibody is an anti-human CD3 monoclonal antibody. 
     
     
       3. The method of  claim 1 , wherein the anti-CD28 antibody is an anti-human CD28 monoclonal antibody. 
     
     
       4. The method of  claim 1 , wherein said solid phase surface is a bead. 
     
     
       5. The method of  claim 4 , wherein the bead is a magnetic immunobead. 
     
     
       6. The method of  claim 1 , wherein said solid phase surface is a tissue culture dish. 
     
     
       7. The method of  claim 1 , wherein the anti-CD3 antibody and the anti-CD28 antibody are immobilized on the solid phase surface via a covalent modification. 
     
     
       8. The method of  claim 1 , wherein the anti-CD3 antibody and the anti-CD28 antibody are immobilized on the solid phase surface via an avidin-biotin complex. 
     
     
       9. The method of  claim 1 , wherein the anti-CD3 antibody and the anti-CD28 antibody are directly immobilized on the solid phase surface. 
     
     
       10. The method of  claim 1 , wherein the T cells are induced to proliferate for at least 3 days. 
     
     
       11. The method of  claim 2 , wherein the T cells are induced to proliferate for at least 7 days. 
     
     
       12. The method of  claim 1 , wherein the T cells are induced to proliferate to about 100-fold the original T cell population. 
     
     
       13. The method of  claim 1 , wherein the T cells are induced to proliferate to about 100,000-fold the original T cell population. 
     
     
       14. The method of  claim 1 , wherein said anti-CD3 antibody and said anti-CD28 antibody are whole antibodies. 
     
     
       15. The method of  claim 1 , wherein the population of T cells is induced to proliferate to sufficient numbers for use in treating an infectious disease. 
     
     
       16. The method of  claim 1 , wherein the population of T cells is induced to proliferate to sufficient numbers for use in treating cancer. 
     
     
       17. A method for inducing a population of T cells to proliferate to sufficient numbers for use in therapy, comprising: 
       a) activating the population of T cells by contacting the T cells in vitro with an anti-CD3 antibody which is immobilized on a solid phase surface; and  
       b) stimulating an accessory molecule on the surface of the T cells in vitro with a stimulatory form of a natural ligand for CD28 selected from the group consisting of B7-1 and B7-2, wherein said stimulatory forrn of a natural ligand for CD28 is immobilized on the same solid phase surface as the anti-CD3 antibody, the activating and stimulating steps thereby inducing proliferation of the T cells to sufficient numbers for use in therapy.  
     
     
       18. The method of  claim 17 , wherein the stimulatory form of a natural ligand for CD28 is B7-1. 
     
     
       19. The method of  claim 17 , wherein the stimulatory form of a natural ligand for CD28 is B7-2. 
     
     
       20. The method of  claim 17 , wherein said solid phase surface is a bead. 
     
     
       21. The method of  claim 20 , wherein the bead is a magnetic immunobead. 
     
     
       22. The method of  claim 17 , wherein said solid phase surface is a tissue culture dish. 
     
     
       23. The method of  claim 17 , wherein the anti-CD3 antibody and the stimulatory form of a natural ligand for anti-CD28 are immobilized on the solid phase surface via a covalent modification. 
     
     
       24. The method of  claim 17 , wherein the anti-CD3 antibody and the stimulatory form of a natural ligand for anti-CD28 are immobilized on the solid phase surface via an avidin-biotin complex. 
     
     
       25. The method of  claim 17 , wherein the anti-CD3 antibody and the stimulatory form of a natural ligand for anti-CD28 are directly immobilized on the solid phase surface. 
     
     
       26. The method of  claim 17 , wherein The T cells are induced to proliferate for at least 3 days. 
     
     
       27. The method of  claim 17 , wherein the T cells are induced to proliferate for at least 7 days. 
     
     
       28. The method of  claim 17 , wherein the T cells are induced to proliferate to about 100-fold the original T cell population. 
     
     
       29. The method of  claim 17 , wherein the T cells are induced to proliferate to about 100,000-fold the original T cell population. 
     
     
       30. The method of  claim 17 , wherein said anti-CD3 antibody is a whole antibody. 
     
     
       31. The method of  claim 17 , wherein the population of T cells is induced to proliferate to sufficient numbers for use in treating an infectious disease. 
     
     
       32. The method of  claim 17 , wherein the population of T cells is induced to proliferate to sufficient numbers for use in treating cancer.